Erc003

The serum levels of AST, ALT, TG, TC, HDL, LDL, and GLU were detected by an automatic biochemical analyzer (AU480, Olympus, Japan). Serum cytokines, including tumor necrosis factor-α (TNF-α) (ERC102a, NeoBioscience, China), monocyte chemotactic protein-1 (MCP-1) (ERC113, NeoBioscience, China), and interleukin- 6 (IL-6) (ERC003, NeoBioscience, China), as well as 25(OH)D₃ (E-EL-0015c, Elabscience, China) were measured by enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions. Liver tissues were homogenized in a working solution and centrifuged at 12000 × g for 20 min. The TG contents (A110-1-1, Jiancheng Bio, China) and FFA contents (A042-2-1, Jiancheng Bio, China) of the liver tissue were detected according to manufacturer’s instructions. The lipid level was normalized with the respective protein concentration.

In common with western blotting, additional tissue was obtained from the ipsilateral basal cortex. The tissues were mechanically homogenized in 0.9% normal saline at 200 mg/mL and centrifuged at 12,000 rpm for 10 min at 4°C. The expression levels of IL-1β (ERC007, Neobioscience, China), IL-6 (ERC003, Neobioscience, China) and TNF-α (ERC102a, Neobioscience, China) expression levels were measured by ELISA, according to the manufacturer’s instructions.